Small-Molecule Analysis Based on DNA Strand Displacement Using a Bacteriorhodopsin Photoelectric Transducer: Taking ATP as an Example

A uniformly oriented purple membrane (PM) monolayer containing photoactive bacteriorhodopsin has recently been applied as a sensitive photoelectric transducer to assay color proteins and microbes quantitatively. This study extends its application to detecting small molecules, using adenosine triphos...

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Bibliographic Details
Published in:Sensors (Basel, Switzerland) Switzerland), 2023-08, Vol.23 (17), p.7453
Main Authors: Chen, Hsiu-Mei, Wang, Wen-Chang, Chen, Hong-Ren
Format: Article
Language:English
Subjects:
Acids
Adenosine triphosphate
Analysis
Antibodies
ATP
Bacteriorhodopsin
Biosensors
Detectors
DNA
Electrodes
Electrolytes
Graphene
Hemoglobin
Immunoassay
Microorganisms
photoelectric biosensor
Proteins
purple membrane
Quantum dots
Reynolds number
Rhodopsin
Sensors
small molecule
strand displacement
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Summary:A uniformly oriented purple membrane (PM) monolayer containing photoactive bacteriorhodopsin has recently been applied as a sensitive photoelectric transducer to assay color proteins and microbes quantitatively. This study extends its application to detecting small molecules, using adenosine triphosphate (ATP) as an example. A reverse detection method is used, which employs AuNPs labeling and specific DNA strand displacement. A PM monolayer-coated electrode is first covalently conjugated with an ATP-specific nucleic acid aptamer and then hybridized with another gold nanoparticle-labeled nucleic acid strand with a sequence that is partially complementary to the ATP aptamer, in order to significantly minimize the photocurrent that is generated by the PM. The resulting ATP-sensing chip restores its photocurrent production in the presence of ATP, and the photocurrent recovers more effectively as the ATP concentration increases. Direct and single-step ATP detection is achieved in 15 min, with detection limits of 5 nM and a dynamic range of 5 nM–0.1 mM. The sensing chip exhibits high selectivity against other ATP analogs and is satisfactorily stable in storage. The ATP-sensing chip is used to assay bacterial populations and achieves a detection limit for Bacillus subtilis and Escherichia coli of 102 and 103 CFU/mL, respectively. The demonstration shows that a variety of small molecules can be simultaneously quantified using PM-based biosensors.
ISSN:1424-8220
1424-8220
DOI:10.3390/s23177453